Development and validation of a sensitive HPLC–ESI-MS/MS method for the direct determination of glucosamine in human plasma

详细信息    查看全文

• 作者：Aldo Roda ; Laura Sabatini ; Anna Barbieri ; Massimo Guardigli ; Marcello Locatelli ; Francesco Saverio Violante ; Lucio C. Rovati ; Stefano Persiani
• 关键词：Bioavailability ; Glucosamine ; HPLC ; Mass spectrometry ; Osteoarthritis
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2006
• 出版时间：21 November 2006
• 年：2006
• 卷：844
• 期：1
• 页码：119-126
• 全文大小：188 K

文摘

A sensitive and specific HPLC–ESI-MS/MS method for the direct determination of glucosamine in human plasma has been developed and validated. Plasma samples were analyzed after a simple, one-step protein precipitation clean-up with trichloroacetic acid using a polymer-based amino high-performance liquid chromatography (HPLC) column and a water/acetonitrile mobile phase elution gradient, with d-[1-¹³C]glucosamine as the internal standard. Detection was performed by mass spectrometry, using an electrospray source and employing multiple reaction monitoring to separately monitor glucosamine and the internal standard. The limit of quantification of the method was 10 ng/ml of glucosamine and the calibration curve showed a good linearity up to 1000 ng/ml. The precision (R.S.D.) and the accuracy (bias) of the method at the limit of quantification were 13.8 and 4.0 % , respectively, and the mean recovery of glucosamine at three concentration levels was 101.6 ± 5.7 % . The method was applied for the determination of glucosamine concentrations in human plasma samples collected from untreated healthy volunteers and, in a separate bioavailability study, to evaluate plasma glucosamine pharmacokinetics profiles after oral administration of crystalline glucosamine sulfate.